Targeting Hydroxybenzoic Acids to Mitochondria as a Strategy to Delay Skin Ageing: An In Vitro Approach.
Targeting antioxidants to mitochondria is considered a promising strategy to prevent cellular senescence and skin ageing. In this study, we investigate whether four hydroxybenzoic acid-based mitochondria-targeted antioxidants (MitoBENs, MB1-4) could be used as potential active ingredients to prevent senescence in skin cells. Firstly, we evaluated the chemical stability, cytotoxicity, genotoxicity and mitochondrial toxicity of all compounds. We followed this by testing the antioxidant protective capacity of the two less toxic compounds on human skin fibroblasts. We then assessed the effects of the best hit on senescence, inflammation and mitochondrial remodeling on a 3D skin cell model, while also testing its mutagenic potential. Cytotoxicity and mitochondrial toxicity rankings were produced: MB3 < MB4 ≃ MB1 < MB2 and MB3 < MB1 < MB4 < MB2, respectively. These results suggest that pyrogallol-based compounds (MB2 and MB4) have lower cytotoxicity. The pyrogallol derivative, MB2, containing a 6-carbon spacer, showed a more potent antioxidant protective activity against hydrogen peroxide cytotoxicity. In a 3D skin cell model, MB2 also decreased transcripts related to senescence. In sum, MB2's biological safety profile, good chemical stability and lack of mutagenicity, combined with its anti-senescence effect, converts MB2 into a good candidate for further development as an active ingredient for skin anti-ageing products.


Nonulcerating bacterial keratitis associated with soft and rigid contact lens wear.
An unusual presentation of contact lens-related bacterial keratitis is that of epithelial nodular infiltrates and stromal inflammation without epithelial ulceration. The authors study the initial diagnosis, clinical features, causative organisms, and outcomes of corneal infections presenting in this manner. The study design was a 20-month retrospective chart review. Five patients with culture-proven bacterial keratitis who had predominantly nodular epithelial lesions were studied. Four infections were associated with soft contact lens wear and one with rigid lens wear. All patients had largely intact epithelium; typical gray-colored epithelial nodules, some with underlying anterior stromal haze; and diffuse, fine, cellular stromal inflammation. Two patients were referred with the tentative diagnosis of Acanthamoeba infection and two as contact lens-related sterile keratitis. Epithelial cultures from three cases yielded Serratia sp., one yielded Corynebacterium, and one Streptococcus pneumoniae. All responded to antibacterial medication; final corrected visual acuity in all cases was 20/30 or better. Bacterial infection associated with contact lens wear can be established within the corneal epithelium without initially producing an ulcer. A wide range of both gram-positive and gram-negative organisms can be involved. Early recognition and treatment appear to result in a favorable outcome.


hSulf-1 inhibits cell proliferation and migration and promotes apoptosis by suppressing stat3 signaling in hepatocellular carcinoma.
Human sulfatase-1 (hSulf-1) has been shown to desulfate cellular heparin sulfate proteoglycans and modulate several growth factors and cytokines. However, hSulf-1 has not been previously shown to mediate the signal transducer and activator of transcription 3 (stat3) signaling pathway, which is known to regulate cell proliferation, motility and apoptosis. The present study investigated the role of hSulf-1 in stat3 signaling in hepatocellular cancer. hSulf-1 expression vector and stat3 small interfering RNA (siRNA) were constructed to control the expression of hSulf-1 and stat3 in HepG2 cells. hSulf-1 was found to inhibit the phosphorylation of stat3 and downregulate its targeted protein. MTT and Transwell chamber assays, as well as Annexin V/propidium iodide double-staining methods, were used to examine the effects of hSulf-1 on stat3-mediated motility, proliferation and apoptosis in HepG2 cells. Transfection with hSulf-1 cDNA and/or stat3 siRNA inhibited cell proliferation and motility, concurrent with G0/G1 and G2/M phase cell cycle arrest and apoptosis. Overall, the results of the current study suggested that hSulf-1 functions as a negative regulator of proliferation and migration and as a positive regulator of apoptosis in hepatocellular carcinoma, at least partly via the downregulation of stat3 signaling.


Pseudolatex preparation using a novel emulsion-diffusion process involving direct displacement of partially water-miscible solvents by distillation.
Pseudolatexes were obtained by a new process based on an emulsification-diffusion technique involving partially water-miscible solvents. The preparation method consisted of emulsifying an organic solution of polymer (saturated with water) in an aqueous solution of a stabilizing agent (saturated with solvent) using conventional stirrers, followed by direct solvent distillation. The technique relies on the rapid displacement of the solvent from the internal into the external phase which thereby provokes polymer aggregation. Nanoparticle formation is believed to occur because rapid solvent diffusion produces regions of local supersaturation near the interface, and nanoparticles are formed due to the ensuing interfacial phase transformations and polymer aggregation that occur in these interfacial domains. Using this method, it was possible to prepare pseudolatexes of biodegradable and non-biodegradable polymers such as poly(D,L-lactic acid) and poly(epsilon-caprolactone), Eudragit E, cellulose acetate phthalate, cellulose acetate trimellitate using ethyl acetate or 2-butanone as partially water-miscible solvents and poly(vinyl alcohol) or poloxamer 407 as stabilizing agent. A transition from nano- to microparticles was observed at high polymer concentrations. At concentrations above 30% w/v of Eudragit E in ethyl acetate or cellulose acetate phthalate in 2-butanone only microparticles were obtained. This behaviour was attributed to decreased transport of polymer molecules into the aqueous phase.


Evidence for formation of DNA repair centers and dose-response nonlinearity in human cells.
The concept of DNA "repair centers" and the meaning of radiation-induced foci (RIF) in human cells have remained controversial. RIFs are characterized by the local recruitment of DNA damage sensing proteins such as p53 binding protein (53BP1). Here, we provide strong evidence for the existence of repair centers. We used live imaging and mathematical fitting of RIF kinetics to show that RIF induction rate increases with increasing radiation dose, whereas the rate at which RIFs disappear decreases. We show that multiple DNA double-strand breaks (DSBs) 1 to 2 μm apart can rapidly cluster into repair centers. Correcting mathematically for the dose dependence of induction/resolution rates, we observe an absolute RIF yield that is surprisingly much smaller at higher doses: 15 RIF/Gy after 2 Gy exposure compared to approximately 64 RIF/Gy after 0.1 Gy. Cumulative RIF counts from time lapse of 53BP1-GFP in human breast cells confirmed these results. The standard model currently in use applies a linear scale, extrapolating cancer risk from high doses to low doses of ionizing radiation. However, our discovery of DSB clustering over such large distances casts considerable doubts on the general assumption that risk to ionizing radiation is proportional to dose, and instead provides a mechanism that could more accurately address risk dose dependency of ionizing radiation.


Calcium carbonate concretions formed by the decomposition of organic matter.
Bacterial decomposition of butterfish and smelts in small sealed jars containing seawater and other solutions, for periods ranging from 65 to 205 days, results in a large increase in concentrations of dissolved bicarbonate, carbonate, and ammonia (plus volatile amines). Accompanying this is a rise in pH and the precipitation of Ca(++) ion from solution. The Ca(++) is not precipitated as CaCO(3) but instead as a mixture of calcium fatty acid salts or soaps with from 14 to 18 carbon atoms. This can be explained by the thermodynamic instability of CaCO(3) relative to Ca soaps in the presence of excess free fatty acid. It is suggested that some ancient CaCO(3) concretions, especially those enclosing fossils of soft-bodied organisms, may have formed rapidly after death in the form of natural Ca soap (adipocere) which was later converted to CaCO(3).


[Interaction between dietary intake and GSTM1 and GSTT1 polymorphisms in head and neck cancer risk: a case-control study in São Paulo, Brazil].
A hospital-based case-control study was conducted to investigate the potential interaction between dietary factors and polymorphisms in phase II metabolic enzymes GSTM1 and GSTT1, associated with head and neck cancer risk. The study included 103 histologically confirmed incident cases and 101 controls. Food intake was estimated with a validated food frequency questionnaire. The gene polymorphisms were evaluated by PCR. Increased risk was observed in the highest tertile of beef consumption in the presence of the GSTM1 (OR = 10.79; 95%CI: 2.17-53.64) and GSTT1 null alleles (OR = 3.41; 95%CI: 0.43-27.21). Assessment of dietary intake considering the ratio between animal product and vegetable consumption showed OR = 2.35 (95%CI: 0.27-19.85) in the intermediate tertile and OR = 3.36 (95%CI: 0.41-27.03) in the highest tertile. The results suggest a possible interaction between meat intake and GSTM1/GSTT1 polymorphisms in modulating the risk of head and neck cancer, influenced by vegetable consumption.


Hospitalizations for acute exacerbations of chronic obstructive pulmonary disease: how you count matters.
ICD-9-CM diagnosis codes are increasingly used to estimate the burden of disease, as well as to evaluate the quality of care and outcomes of various conditions. Acute exacerbations of COPD (AE-COPD) are common and associated with substantial health and financial burden in the U.S. Whether published algorithms that employ different combinations of ICD-9-CM codes to identify patients hospitalized for AE-COPD yield similar or different estimates of disease burden is unclear. In this study, the Nationwide Inpatient Sample from years 2000-2006 was used to identify and compare the number of hospitalizations, healthcare utilization, and outcomes for patients hospitalized for AE-COPD in the U.S. AE-COPD was identified using five different published ICD-9-CM algorithms. Estimates of the annual number of hospitalizations for AE-COPD in the U.S. varied more than 2-fold (e.g., 421,000 to 870,000 in 2006). Outcomes and healthcare utilization of patients hospitalized for AE-COPD varied substantially, depending on the algorithm used (e.g., in-hospital mortality 2.0% to 5.1%, total hospital days 2.0 to 5.1 million in 2006). Observed trends in the number of hospitalizations over the 7-year period varied depending on which algorithm was used. In conclusion, the estimated health burden and trends in hospitalizations for AE-COPD in the United States differ, depending on which ICD-9-CM algorithm is used. To improve our understanding of the burden of AE-COPD and to ensure that quality of care initiatives are not misdirected, a validated approach to identifying patients hospitalized for AE-COPD is needed.


The role of tricuspid valve surgery in the late management of tetralogy of fallot: collective review.
While surgical repair of tetralogy of Fallot (TOF) is generally associated with good early outcomes, late complications affect long-term survival and may require reoperation. Pulmonary regurgitation (PR) and tricuspid regurgitation (TR) may increase the risk of arrhythmias, reduced cardiac function, and sudden death. Tricuspid valve function can be compromised secondarily in the setting of PR or directly as a result of injury or alteration of the valve related to the original TOF repair. This article reviews the etiologic mechanisms, pathophysiological implications, and surgical interventions for TR. Effective management following TOF repair requires consideration of TR to optimize late outcomes.


The effect of indomethacin of HMG-HCG induced ovulation in the phesus monkey.
The investigation was designed to study the influence of indomethacin on gonadotropin induced ovulation in the rhesus monkey. Six mature female monkeys were treated with HMG-HCG for at least 2 control ovulatory cycles at dosage levels adjusted to induced ovulation while avoiding superovulation. Ovulation was confirmed by observation of the ovaries for fresh ovulation points at laparotomy. Following establishment of an appropriate dosage schedule, treatment was begun with indomethacin (5 mg/kg/day) starting 5 days prior to HCG and continuing to the time of laparotomy. In a second treatment cycle, indomethacin was administered at a dose of 5 mg/kg b.i.d. together with the established dose of HMG-HCG. Ovarian inspection was carried out as in the control cycles. Venous blood was obtained on treatment days 4, 7, 10 and 11 for determination of serum estrone, estradiol and progesterone. Indomethacin administration resulted in ovulation inhibition at a dose of 10 mg/kg/day when ovulation inducing doses of gonadotropins were administered. Peripheral blood steroid levels suggest that follicle maturation and estrogen production was unimparied by indomethacin. These findings indicate that the ovarian synthesis of prostaglandin may be essential in the process of ovulation.


Effective reduction of nitrophenols and colorimetric detection of Pb(ii) ions by Siraitia grosvenorii fruit extract capped gold nanoparticles.
This study presents a simple and green approach for the synthesis of Siraitia grosvenorii fruit extract capped gold nanoparticles (SG-AuNPs). The SG-AuNPs samples prepared under the optimized conditions were characterized by various techniques (UV-Vis, XRD, FTIR, HR-TEM, EDX, DLS). The biosynthesized nanoparticles were then studied for the reduction of 2-nitrophenol (2-NP) and 3-nitrophenols (3-NP) and for colorimetric detection of Pb2+ ions. The characterization results revealed that the crystals of SG-AuNPs were spherical with an average size of 7.5 nm. The FTIR and DLS analyses proved the presence of the biomolecule layer around AuNPs, which played an important role in stabilizing the nanoparticles. The SG-AuNPs showed excellent catalytic activity in the reduction of 3-NP and 2-NP, achieving complete conversion within 14 min. The catalytic process was endothermic and followed pseudo-first-order kinetics. The activation energy was determined to be 10.64 and 26.53 kJ mol-1 for 2-NP and 3-NP, respectively. SG-AuNPs maintained high catalytic performance after five recycles. The fabricated material was also found to be highly sensitive and selective to Pb2+ ions with the detection limit of 0.018 μM in a linear range of 0-1000 μM. The practicality of the material was validated through the analyses of Pb2+ in mimic pond water samples. The developed nanoparticles could find tremendous applications in environmental monitoring.


Acute chemokine response in the blood and cerebrospinal fluid of children with enterovirus 71-associated brainstem encephalitis.
Brainstem encephalitis (BE) is a serious neurological complication of enterovirus 71 (EV71) infection. The present study was designed to determine the characteristics of the chemokine response in the blood and cerebrospinal fluid (CSF) of patients with EV71-associated BE. Thirty-one patients with BE were studied. They consisted of 12 with uncomplicated BE, 9 with autonomic nervous system (ANS) dysregulation, and 10 with pulmonary edema (PE); 13 healthy control subjects were also studied. Plasma and CSF concentrations of various chemokines were determined by a particle-based flow cytometry immunoassay. Plasma levels of interferon (IFN)-gamma-induced protein (IP)-10, monocyte chemoattractant protein (MCP)-1, monokine induced by IFN-gamma (MIG), and interleukin (IL)-8 were significantly higher in patients with PE than in those with uncomplicated BE. CSF levels of MIG were significantly higher in patients with PE than in those with uncomplicated BE and ANS dysregulation. The ratios of mean CSF to plasma levels for MCP-1 and IL-8 were highest in patients with uncomplicated BE and tended to fall with increasing severity of the disease. Overexpression of the chemokine cascade in the central nervous system compartment appears to play an important role in the elicitation of the immune response to EV71. The chemokine CSF to plasma ratios suggest that IL-8, IP-10, MCP-1, and possibly MIG-but not RANTES-are synthesized in the brain in response to encephalitis.


Spotted fever group rickettsiae in ticks, Germany.
To explore increased risk for human Rickettsia spp. infection in Germany, we investigated recreational areas and renatured brown coal surface-mining sites (also used for recreation) for the presence of spotted fever group rickettsiae in ticks. R. raoultii (56.7%), R. slovaca (13.3%), and R. helvetica (>13.4%) were detected in the respective tick species.


Giant hemorrhagic trochanteric bursitis mimicking a high-grade soft tissue sarcoma: report of two cases.
The giant haemorrhagic bursitis of the hip joint is a rare clinical condition that requires evidence-based guidelines for adequate diagnosis and management. Usually, this pathology requires conservative treatment; however, when abnormal size or clinical symptoms of compression of the surrounding noble structures are reported, an accurate differential diagnosis is required, in order to exclude other malignant conditions that can be included into differential diagnosis, and a surgical approach should be considered. The purpose of this work is to provide an appropriate description of the diagnostic and therapeutic path, providing an accurate analysis of the possible differential diagnoses. We report 2 cases of symptomatic haemorrhagic bursitis of the hip joint, confirmed by histological investigation. In both cases, the patients complained a peripheral nerve deficit of a single limb: one patient presented paresthesia of lateral femoral cutaneous nerve while the second peripheral edema due to compression of the proximal venous and lymphatic circulation. Both cases were successfully managed by complete surgical excision of the mass, with no recurrence. There were no major complications, but in first case the nerve deficit was permanent. Giant hemorrhagic trochanteric bursitis is a rare condition, but it should be included in the differential diagnosis of soft tissue masses arising from the hip joint. Due to the rarity of this entity, a cautious exclusion process of all plausible differential diagnosis must be undertaken, in order to not miss the possibility of soft-tissue tumors, primarily malignant high-grade sarcomas.


Pleural malignancies.
Pleural malignancies, primary or metastatic, portend a grim prognosis. In addition to the serious oncologic implications of a pleural malignancy, these tumors can be highly symptomatic. A malignant pleural effusion can cause dyspnea, secondary to lung compression, or even tension physiology from a hydrothorax under pressure. The need to palliate these effusions is a seemingly straightforward clinical scenario, but with nuances that can result in disastrous complications for the patient if not attended to appropriately. Solid pleural malignancies can cause great pain from chest wall invasion or can cause a myriad of morbid symptoms because of the invasion of thoracic structures, such as the heart, lungs, or esophagus. This article reviews pleural malignancies, the purely palliative treatments, and the treatments that are performed with definitive (curative) intent.


Comprehensive Quantitative Profiling of Tau and Phosphorylated Tau Peptides in Cerebrospinal Fluid by Mass Spectrometry Provides New Biomarker Candidates.
Aberrant tau phosphorylation is a hallmark in Alzheimer's disease (AD), believed to promote formation of paired helical filaments, the main constituent of neurofibrillary tangles in the brain. While cerebrospinal fluid (CSF) levels of total tau and tau phosphorylated at threonine residue 181 (pThr181) are established core biomarkers for AD, the value of alternative phosphorylation sites, which may have more direct relevance to pathology, for early diagnosis is not yet known, largely due to their low levels in CSF and lack of standardized detection methods. To overcome sensitivity limitations for analysis of phosphorylated tau in CSF, we have applied an innovative mass spectrometry (MS) workflow, TMTcalibratortrademark, to enrich and enhance the detection of phosphoproteome components of AD brain tissue in CSF, and enable the quantitation of these analytes. We aimed to identify which tau species present in the AD brain are also detectable in CSF and which, if any, are differentially regulated with disease. Over 75% coverage of full-length (2N4R) tau was detected in the CSF with 47 phosphopeptides covering 31 different phosphorylation sites. Of these, 11 phosphopeptides were upregulated by at least 40%, along with an overall increase in tau levels in the CSF of AD patients relative to controls. Use of the TMTcalibratortrademark workflow dramatically improved our ability to detect tau-derived peptides that are directly related to human AD pathology. Further validation of regulated tau peptides as early biomarkers of AD is warranted and is currently being undertaken.


Marine Sulfated Polysaccharides as Promising Antiviral Agents: A Comprehensive Report and Modeling Study Focusing on SARS CoV-2.
SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) is a novel coronavirus strain that emerged at the end of 2019, causing millions of deaths so far. Despite enormous efforts being made through various drug discovery campaigns, there is still a desperate need for treatments with high efficacy and selectivity. Recently, marine sulfated polysaccharides (MSPs) have earned significant attention and are widely examined against many viral infections. This article attempted to produce a comprehensive report about MSPs from different marine sources alongside their antiviral effects against various viral species covering the last 25 years of research articles. Additionally, these reported MSPs were subjected to molecular docking and dynamic simulation experiments to ascertain potential interactions with both the receptor-binding domain (RBD) of SARS CoV-2's spike protein (S-protein) and human angiotensin-converting enzyme-2 (ACE2). The possible binding sites on both S-protein's RBD and ACE2 were determined based on how they bind to heparin, which has been reported to exhibit significant antiviral activity against SARS CoV-2 through binding to RBD, preventing the virus from affecting ACE2. Moreover, our modeling results illustrate that heparin can also bind to and block ACE2, acting as a competitor and protective agent against SARS CoV-2 infection. Nine of the investigated MSPs candidates exhibited promising results, taking into consideration the newly emerged SARS CoV-2 variants, of which five were not previously reported to exert antiviral activity against SARS CoV-2, including sulfated galactofucan (1), sulfated polymannuroguluronate (SPMG) (2), sulfated mannan (3), sulfated heterorhamnan (8), and chondroitin sulfate E (CS-E) (9). These results shed light on the importance of sulfated polysaccharides as potential SARS-CoV-2 inhibitors.


Induction of partial mitosis in BHK cells by 2-aminopurine.
The protein kinase inhibitor 2-aminopurine (2-AP) inhibits a subset of mitotic events in BHK cells. In the presence of the drug, these cells form a bipolar spindle in mitosis, but chromatin fails to generate functioning chromosomes. Cells in 2-AP progress through a partial mitosis, in which there is no observable metaphase, anaphase or telophase events. After 12 h of exposure to 2-AP the chromatin in mitotic cells fails to condense into discrete chromosomes, and is displaced by the spindle to form 'binucleate' cells and cells containing abnormally shaped nuclei in the subsequent interphase. Other mitotic modifications of nuclei, such as nucleolar and nuclear lamina disassembly, occur normally. Centromeres in these nuclei do not become engaged in the spindle, but instead show either no association or a lateral arrangement around the spindle. Cells treated with 2-AP are not arrested in mitosis. Therefore, mitotic exit is not inhibited by the failure of these cells to progress through the latter stages of mitosis. Further, nocodazole-arrested cells quickly exit mitotic arrest when 2-AP is added. We conclude that 2-AP interferes with a specific subset of mitotic events, and that it allows cells to overcome check-points that require spindle function for mitotic progression.


Serum endotrophin identifies optimal responders to PPARγ agonists in type 2 diabetes.
The treatment of type 2 diabetes with full peroxisome proliferator-activated receptor gamma (PPARγ) agonists improves insulin sensitivity, but is associated with weight gain, heart failure, peripheral oedema and bone loss. Endotrophin, the C-terminal fragment of the α3 chain of procollagen type VI (also called Pro-C6), is involved in both adipose tissue matrix remodelling and metabolic control. We established a serum assay for endotrophin to assess if this novel adipokine could identify type 2 diabetic patients who respond optimally to PPARγ agonists, improving the risk-to-benefit ratio. The BALLET trial (NCT00515632) compared the glucose-lowering effects and safety of the partial PPARγ agonist balaglitazone with those of pioglitazone in individuals with type 2 diabetes on stable insulin therapy. The per protocol population (n = 297) was stratified into tertiles based on baseline endotrophin levels. Participants were followed-up after 26 weeks, after which correlational analysis was carried out between endotrophin levels and measures of glucose control. This is a secondary post hoc analysis. Endotrophin was significantly associated with therapeutic response to balaglitazone and pioglitazone. At week 26, only individuals in the upper two tertiles showed significant reductions in HbA1c and fasting serum glucose compared with baseline. The OR for a 1% and a 0.5% reduction in HbA1c for individuals in the upper two tertiles were 3.83 (95% CI 1.62, 9.04) p < 0.01, and 3.85 (95% CI 1.94, 7.61) p < 0.001, respectively. Endotrophin levels correlated with adipose tissue mass, insulin resistance and fatty liver index. Notably, PPARγ-associated adverse effects, such as moderate-to-severe lower extremity oedema, only occurred in the lower tertile. Elevated endotrophin serum levels predict response to two insulin sensitisers and reduce the risk of associated adverse effects, thereby, identifying patients with type 2 diabetes who may profit from PPARγ agonist treatment.


Identification of genes regulating migration and invasion using a new model of metastatic prostate cancer.
Understanding the complex, multistep process of metastasis remains a major challenge in cancer research. Metastasis models can reveal insights in tumor development and progression and provide tools to test new intervention strategies. To develop a new cancer metastasis model, we used DU145 human prostate cancer cells and performed repeated rounds of orthotopic prostate injection and selection of subsequent lymph node metastases. Tumor growth, metastasis, cell migration and invasion were analyzed. Microarray analysis was used to identify cell migration- and cancer-related genes correlating with metastasis. Selected genes were silenced using siRNA, and their roles in cell migration and invasion were determined in transwell migration and Matrigel invasion assays. Our in vivo cycling strategy created cell lines with dramatically increased tumorigenesis and increased ability to colonize lymph nodes (DU145LN1-LN4). Prostate tumor xenografts displayed increased vascularization, enlarged podoplanin-positive lymphatic vessels and invasive margins. Microarray analysis revealed gene expression profiles that correlated with metastatic potential. Using gene network analysis we selected 3 significantly upregulated cell movement and cancer related genes for further analysis: EPCAM (epithelial cell adhesion molecule), ITGB4 (integrin β4) and PLAU (urokinase-type plasminogen activator (uPA)). These genes all showed increased protein expression in the more metastatic DU145-LN4 cells compared to the parental DU145. SiRNA knockdown of EpCAM, integrin-β4 or uPA all significantly reduced cell migration in DU145-LN4 cells. In contrast, only uPA siRNA inhibited cell invasion into Matrigel. This role of uPA in cell invasion was confirmed using the uPA inhibitors, amiloride and UK122. Our approach has identified genes required for the migration and invasion of metastatic tumor cells, and we propose that our new in vivo model system will be a powerful tool to interrogate the metastatic cascade in prostate cancer.


mGluR2 versus mGluR3 Metabotropic Glutamate Receptors in Primate Dorsolateral Prefrontal Cortex: Postsynaptic mGluR3 Strengthen Working Memory Networks.
The newly evolved circuits in layer III of primate dorsolateral prefrontal cortex (dlPFC) generate the neural representations that subserve working memory. These circuits are weakened by increased cAMP-K+ channel signaling, and are a focus of pathology in schizophrenia, aging, and Alzheimer's disease. Cognitive deficits in these disorders are increasingly associated with insults to mGluR3 metabotropic glutamate receptors, while reductions in mGluR2 appear protective. This has been perplexing, as mGluR3 has been considered glial receptors, and mGluR2 and mGluR3 have been thought to have similar functions, reducing glutamate transmission. We have discovered that, in addition to their astrocytic expression, mGluR3 is concentrated postsynaptically in spine synapses of layer III dlPFC, positioned to strengthen connectivity by inhibiting postsynaptic cAMP-K+ channel actions. In contrast, mGluR2 is principally presynaptic as expected, with only a minor postsynaptic component. Functionally, increase in the endogenous mGluR3 agonist, N-acetylaspartylglutamate, markedly enhanced dlPFC Delay cell firing during a working memory task via inhibition of cAMP signaling, while the mGluR2 positive allosteric modulator, BINA, produced an inverted-U dose-response on dlPFC Delay cell firing and working memory performance. These data illuminate why insults to mGluR3 would erode cognitive abilities, and support mGluR3 as a novel therapeutic target for higher cognitive disorders.


On Aggregation of Unsupervised Deep Binary Descriptor with Weak Bits.
Despite the thrilling success achieved by existing binary descriptors, most of them are still in the mire of three limitations: 1) vulnerable to the geometric transformations; 2) incapable of preserving the manifold structure when learning binary codes; 3) NO guarantee to find the true match if multiple candidates happen to have the same Hamming distance to a given query. All these together make the binary descriptor less effective, given large-scale visual recognition tasks. In this paper, we propose a novel learning-based feature descriptor, namely Unsupervised Deep Binary Descriptor (UDBD), which learns transformation invariant binary descriptors via projecting the original data and their transformed sets into a joint binary space. Moreover, we involve a ℓ2,1-norm loss term in the binary embedding process to gain simultaneously the robustness against data noises and less probability of mistakenly flipping bits of the binary descriptor, on top of it, a graph constraint is used to preserve the original manifold structure in the binary space. Furthermore, a weak bit mechanism is adopted to find the real match from candidates sharing the same minimum Hamming distance, thus enhancing matching performance. Extensive experimental results on public datasets show the superiority of UDBD in terms of matching and retrieval accuracy over state-of-the-arts.


Intimate partner violence exposure during infancy and social functioning in middle childhood: An Australian mother and child cohort study.
Social functioning of children with experiences of intimate partner violence (IPV) between caregivers in early childhood has received less attention than emotional-behavioral outcomes. Drawing on data from 1507 ten-year-old Australian-born children and their mothers participating in a community-based longitudinal study, this study examined the associations between IPV exposure during infancy and social development during middle childhood. IPV during the first 12 months of life was associated with lower social skills, higher peer problems, and peer victimization at age 10 years, while accounting for concurrent IPV. This study provides evidence for the long-term impacts of early-life IPV exposure on children's social functioning, and the importance of prevention and early intervention programs focused on social development following experiences of IPV.


The Protein Interactome of Streptococcus pneumoniae and Bacterial Meta-interactomes Improve Function Predictions.
The functions of roughly a third of all proteins in Streptococcus pneumoniae, a significant human-pathogenic bacterium, are unknown. Using a yeast two-hybrid approach, we have determined more than 2,000 novel protein interactions in this organism. We augmented this network with meta-interactome data that we defined as the pool of all interactions between evolutionarily conserved proteins in other bacteria. We found that such interactions significantly improved our ability to predict a protein's function, allowing us to provide functional predictions for 299 S. pneumoniae proteins with previously unknown functions. IMPORTANCE Identification of protein interactions in bacterial species can help define the individual roles that proteins play in cellular pathways and pathogenesis. Very few protein interactions have been identified for the important human pathogen S. pneumoniae. We used an experimental approach to identify over 2,000 new protein interactions for S. pneumoniae, the most extensive interactome data for this bacterium to date. To predict protein function, we used our interactome data augmented with interactions from other closely related bacteria. The combination of the experimental data and meta-interactome data significantly improved the prediction results, allowing us to assign possible functions to a large number of poorly characterized proteins.


[Matrix-producing breast carcinoma with myoepithelial differentiation--description of 11 cases and review of literature aimed at histogenesis and differential diagnosis].
Presentation of variable morphology of the so-called matrix-producing carcinoma, the rare variant of metaplastic breast carcinoma. Establish the extent of myoepithelial phenotype by immunohistochemistry and assess behaviour of the lesion. Review of clinicopathologic data of 11 cases. Private Biopsy Lab s.r.o., Pilsen and Sikl's Department of Pathology, Charles University and Faculty Hospital, Pilsen. Included are eleven cases from Breast Registry, Biopsy Lab s.r.o., Pilsen and Pathology Department, Faculty Hospital in Hradec Králové and Martin and from Biopsy Lab in Horovice. Analyzed were morphology, stage, grade and follow up. Myoepithelial phenotype was demonstrated immunohistochemically by LSAB+ system (DAKO) and seven conventional myoepithelial markers (GFAP, aktin S, calponin, cytokeratin 14, p63, CD10 and P-cadherin) were used. Oncogene p53 and Her2/neu were also detected. Matrix-producing carcinoma is an extremely rare type of metaplastic carcinoma of the breast, where high-grade epithelial component continually merges with heterologous mesenchymal chondroid component without overt spindle cell sarcomatoid pattern in between. There were all women at the age of 32 to 86-years (average 54 years, median 52 years) and the maximum tumor diameter ranged from 18 to 60 mm (average 32 mm). The axillary LN was positive in six cases in the time of diagnosis. The follow up available in 9 women ranged from three months to ten years (average 24 months); dissemination of the disease was observed in three cases; 2 of these patients died of disease after three and ten years, respectively. The heterologous mesenchymal chondroid component was produced in two different ways; one displayed typical structure of low-grade hyaline cartilage, in the second one the epithelial tumorous cells were embedded in the homogenous eosinophilic extracellular matrix giving appearance of chondroid aura. Metaplastic component constituted 5-75% of the tumor volume. Immunohistochemical evidence of myoepithelial differentiation in all neoplasms was demonstrated, with at least two conventional myoepithelial markers (actin S, calponin, GFAP, CK14, CD10, p63 and P-cadherin) being positive in every case. Expression of p53 was identified in six cases, all tumors were Her2/neu negative. Histologically the metastases were formed either by carcinoma cells only, or more frequently, they replicated the structure of primary lesion. As a rare entity, matrix-producing carcinoma of breast displaying myoepithelial phenotype, deserves separate position in tumor classification. It differs from conventional myoepithelial carcinoma and from heterologous metaplastic carcinoma, where the matrix emanate from undifferentiated sarcomatous tissue, but precise histogenesis in not clear yet. This is a very aggressive lesion and our findings show that previous reports of indolent behaviour were preliminary. Spectrum of differential diagnosis, namely benign mixed tumor, phylloid tumor, primary breast sarcoma and colloid carcinoma warrant clear knowledge about this unique entity.


[Transosseous osteosynthesis in fractures of bones of the elbow joint].
The experience with the treatment of 64 patients with fresh fractures of the elbow joint bones by the method of percutaneous osteosynthesis with the Ilizarov apparatus has shown its high efficiency in the treatment of such injuries.


Paleomobility in the Tiwanaku diaspora: biogeochemical analyses at Rio Muerto, Moquegua, Peru.
Paleomobility has been a key element in the study of the expansion of ancient states and empires, including the Tiwanaku polity of the South Central Andes (AD 500-1000). We present radiogenic strontium and oxygen isotope data from human burials from three cemeteries in the Tiwanaku-affiliated Middle Horizon archaeological site complex of Rio Muerto in the Moquegua Valley of southern Peru. At Rio Muerto, archaeological human enamel and bone values range from (87) Sr/(86) Sr = 0.70657-0.72018, with a mean of (87) Sr/(86) Sr = 0.70804 ± 0.00207 (1σ, n = 55). For the subset of samples analyzed for oxygen isotope values (n = 48), the data ranges from δ(18) Ocarbonate(VSMOW) = +18.1 to +27.0‰. When contextualized with other lines of archaeological evidence, we interpret these data as evidence for an archaeological population in which the majority of individuals had "local" origins, and were likely second-generation, or more, immigrants from the Tiwanaku heartland in the altiplano. Based on detailed life history data, we argue a smaller number of individuals came at different ages from various regions within the Tiwanaku polity. We consider whether these individuals with isotopic values consistent with "nonlocal" geographic origins could represent first-generation migrants, marriage exchange partners, or occupationally mobile herders, traders or other travelers. By combining isotopic life history studies with mortuary treatment data, we use a person-centered migration history approach to state integration and expansion. Isotopic analyses of paleomobility at the Rio Muerto site complex contribute to the role of diversity in ancient states by demonstrating the range of geographic origins rather than simply colonists from the Lake Titicaca Basin.


Maximizing the efficacy of SAGE analysis identifies novel transcripts in Arabidopsis.
The efficacy of using Serial Analysis of Gene Expression (SAGE) to analyze the transcriptome of the model dicotyledonous plant Arabidopsis was assessed. We describe an iterative tag-to-gene matching process that exploits the availability of the whole genome sequence of Arabidopsis. The expression patterns of 98% of the annotated Arabidopsis genes could theoretically be evaluated through SAGE and using an iterative matching process 79% could be identified by a tag found at a unique site in the genome. A total of 145,170 reliable experimental tags from two Arabidopsis leaf tissue SAGE libraries were analyzed, of which 29,632 were distinct. The majority (93%) of the 12,988 experimental tags observed greater than once could be matched within the Arabidopsis genome. However, only 78% were matched to a single locus within the genome, reflecting the complexities associated with working in a highly duplicated genome. In addition to a comprehensive assessment of gene expression in Arabidopsis leaf tissue, we describe evidence of transcription from pseudo-genes as well as evidence of alternative mRNA processing and anti-sense transcription. This collection of experimental SAGE tags could be exploited to assist in the on-going annotation of the Arabidopsis genome.


Design and analysis issues in cardiac arrhythmia trials: insights from the Detect Supraventricular Tachycardia Trial.
The Detect Supraventricular Tachycardia (Detect SVT) trial was a randomized single blind study in subjects receiving a dual chamber implantable cardioverter defibrillator (ICD). The primary goal was to evaluate whether dual chamber enhancements result in greater discrimination between supraventricular tachycardia (SVT) and ventricular episodes when compared to ventricular-only enhancements. The purpose of this report is to describe our experiences from an analytic perspective, including overall study design, the value of blinded episode review, and the statistical analysis plan. The Detect SVT study was a parallel group study, with blinded central review of all arrhythmia episodes occurring over a six-month period. The primary response was the proportion of inappropriate detections (SVT episodes that are inappropriately classified as ventricular by the device). A parallel group design was essential as unplanned crossovers were very differential. The blinded review of episodes was a major use of resources, with over 2300 episodes observed. This review did not materially alter the findings, however, the avoidance of any perceived bias was a major strength. The value of collecting large numbers of episodes on a single subject was found to be minimal. A Kaplan-Meier analysis of the number of SVT episodes until first inappropriate detection was found to add additional information. The recommendations reported here relate to cardiac arrhythmia trials, or other studies with very similar endpoints. These observations are based on the results of a single trial and should be confirmed in other studies. The parallel group design was essential, as was accounting for within-subject correlation in the analysis. The blinded review process was also important, but significant resources could be saved by closing-out episode collection after a fixed number of episodes. Lessons learned from the Detect SVT trial may prove useful to others designing trials in this active research area.


Delivering the Lee Silverman Voice Treatment (LSVT) by web camera: a feasibility study.
Speech disorders are a feature of Parkinson's disease, typically worsening as the disease progresses. The Lee Silverman Voice Treatment (LSVT) was developed to address these difficulties. It targets vocal loudness as a means of increasing vocal effort and improving coordination across the subsystems of speech. Currently LSVT is not widely available, and there are practical difficulties associated with the delivery of an intensive treatment in an environment of resource constraints in the National Health Service (NHS). Delivery of LSVT over the World-Wide Web may address some of these difficulties. A feasibility study is reported in which three individuals with speech disorders resulting from Parkinson's disease received LSVT over the Internet, using broadband connection and a web camera. Participants were seen face to face for every fourth session in order to build a personal relationship, measure vocal sound pressure level (SPL) during treatment, and to review and prepare homework tasks. All other sessions were delivered over the Internet. Broadly similar treatment gains were found between individuals treated over the Internet and those treated face to face. Gains were maintained or improved at an assessment two months after the treatment. This study demonstrates that delivery of LSVT over the Internet is feasible, and that a larger trial would be appropriate to establish cost and treatment effectiveness.


Eyepass Glaucoma Implant in Open-Angle Glaucoma After Failed Conventional Medical Therapy: Clinical Results of a 5-Year-Follow-up.
The purpose of the study was to evaluate the long-term safety and intraocular pressure (IOP) lowering effect of the Eyepass glaucoma implant (GMP Vision Solutions, Inc.). The prospective study included 15 patients (16 eyes) with primary open-angle glaucoma who underwent an implantation of the Y-shaped Eyepass glaucoma implant. This shunt diverts aqueous from the anterior chamber directly into Schlemm's canal to increase outflow and to lower the IOP. IOP, visual acuity, potential complications and the number of antiglaucomatous medications were monitored over a period of 5 years. The implant was successfully inserted in 14 of 16 eyes. Mean IOP was reduced from 26.4±8.1 mm Hg (SD) to 16.4±5.3 mm Hg (P=0.032) at the end of the follow-up. Mean number of antiglaucomatous medications dropped from 2.1±1.2 (SD) to 0.9±1.2 (SD). In 5 cases, no pressure-lowering medications were necessary 5 years after surgery. Mean best-corrected visual acuity did not change significantly (P>0.05). In all cases, filtering blebs were observed and sustained using antimetabolites. The most common complication was temporary ocular hypotony. Two patients required a revision surgery due to implant malposition. The Eyepass glaucoma implant seems to be a safe and effective treatment option for patients with primary open-angle glaucoma. The use of this device resulted in a significant decrease of IOP.


Biotransformation of phytoestrogens from soy in enzymatically characterized liver microsomes and primary hepatocytes of Atlantic salmon.
Efficient aquaculture is depending on sustainable protein sources. The shortage in marine raw materials has initiated a shift to "green aquafeeds" based on staple ingredients such as soy and wheat. Plant-based diets entail new challenges regarding fish health, product quality and consumer risks due to the possible presence of chemical contaminants, natural toxins and bioactive compounds like phytoestrogens. Daidzein (DAI), genistein (GEN) and glycitein (GLY) are major soy isoflavones with considerable estrogenic activities, potentially interfering with the piscine endocrine system and affecting consumers after carry-over. In this context, information on isoflavone biotransformation in fish is crucial for risk evaluation. We have therefore isolated hepatic fractions of Atlantic salmon (Salmo salar), the most important species in Norwegian aquaculture, and used them to study isoflavone elimination and metabolite formation. The salmon liver microsomes and primary hepatocytes were characterized with respect to phase I cytochrome P450 (CYP) and phase II uridine-diphosphate-glucuronosyltransferase (UGT) enzyme activities using specific probe substrates, which allowed comparison to results in other species. DAI, GEN and GLY were effectively cleared by UGT. Based on the measurement of exact masses, fragmentation patterns, and retention times in liquid chromatography high-resolution mass spectrometry, we preliminarily identified the 7-O-glucuronides as the main metabolites in salmon, possibly produced by UGT1A1 and UGT1A9-like activities. In contrast, the production of oxidative metabolites by CYP was insignificant. Under optimized assay conditions, only small amounts of mono-hydroxylated DAI were detectable. These findings suggested that bioaccumulation of phytoestrogens in farmed salmon and consumer risks from soy-containing aquafeeds are unlikely.


Calbindin-immunoreactive neurons in the reticular formation of the rat brainstem: catecholamine content and spinal projections.
Calbindin-D28k (calbindin) is a calcium-binding protein that is distributed widely in the rat brain. The localisation of calbindin immunoreactivity in the medulla oblongata and its colocalisation with adrenaline-synthesising neurons [phenylethanolamine-N-methyltransferase-immunoreactive (PNMT-IR)] was examined (Granata and Chang [1994] Brain Res. 645:265-277). However, detailed information about the distribution of calbindin-IR neurons in the reticular formation of the medulla oblongata in particular is lacking. In this report, the authors address this issue with an emphasis on the quantitation of calbindin-IR neurons, catecholamine neurons [tyrosine hydroxylase (TH)-IR, or PNMT-IR], and spinally projecting neurons in the ventral brainstem. Rats received injections of the retrograde tracing agent cholera toxin B (CTB) into the thoracic spinal cord or into the superior cervical ganglion. Immunocytochemistry was used to reveal calbindin, TH, PNMT, and CTB immunoreactivity. Ten calbindin-IR cell groups were identified within the pontomedullary reticular formation. Seven previously undescribed but distinct clusters of calbindin-IR neurons were found. Within the ventral pons, a population of calbindin-IR neurons occurred dorsal but adjacent to the A5 cell group. These calbindin-IR neurons did not contain either TH or PNMT immunoreactivity, and few if any of these neurons projected to the spinal cord. A distinct group of calbindin-IR neurons was present in the ventral medulla. Seventy-five percent of these calbindin-IR neurons contained TH immunoreactivity, 45% contained PNMT immunoreactivity, and 21% were spinally projecting neurons. Spinally projecting, calbindin-IR neurons were a subpopulation of PNMT-IR cells. In the caudal ventral medulla, no TH-IR or PNMT-IR cells were calbindin-IR. In the intermediolateral cell column, close appositions of calbindin-IR terminals on identified sympathetic preganglionic neurons as well as calbindin-IR synapses indicated that these neurons may affect directly the sympathetic outflow. The results demonstrate for the first time the existence of a new subpopulation of spinally projecting, PNMT-IR neurons in the rostral ventrolateral medulla.


Lysine 4 of histone H3.3 is required for embryonic stem cell differentiation, histone enrichment at regulatory regions and transcription accuracy.
Mutations in enzymes that modify histone H3 at lysine 4 (H3K4) or lysine 36 (H3K36) have been linked to human disease, yet the role of these residues in mammals is unclear. We mutated K4 or K36 to alanine in the histone variant H3.3 and showed that the K4A mutation in mouse embryonic stem cells (ESCs) impaired differentiation and induced widespread gene expression changes. K4A resulted in substantial H3.3 depletion, especially at ESC promoters; it was accompanied by reduced remodeler binding and increased RNA polymerase II (Pol II) activity. Regulatory regions depleted of H3.3K4A showed histone modification alterations and changes in enhancer activity that correlated with gene expression. In contrast, the K36A mutation did not alter H3.3 deposition and affected gene expression at the later stages of differentiation. Thus, H3K4 is required for nucleosome deposition, histone turnover and chromatin remodeler binding at regulatory regions, where tight regulation of Pol II activity is necessary for proper ESC differentiation.


Risk factors for incident chronic insomnia: a general population prospective study.
The few population-based, prospective studies that have examined risk factors of incident insomnia were limited by small sample size, short follow-up, and lack of data on medical disorders or polysomnography. We prospectively examined the associations between demographics, behavioral factors, psychiatric and medical disorders, and polysomnography with incident chronic insomnia. From a random, general population sample of 1741 individuals of the adult Penn State Sleep Cohort, 1395 were followed-up after 7.5 years. Only subjects without chronic insomnia at baseline (n = 1246) were included in this study. Structured medical and psychiatric history, personality testing, and 8-h polysomnography were obtained at baseline. Structured sleep history was obtained at baseline and follow-up. Incidence of chronic insomnia was 9.3%, with a higher incidence in women (12.9%) than in men (6.2%). Younger age (20-35 years), non-white ethnicity, and obesity increased the risk of chronic insomnia. Poor sleep and mental health were stronger predictors of incident chronic insomnia compared to physical health. Higher scores in MMPI-2, indicating maladaptive personality traits, and excessive use of coffee at baseline predicted incident chronic insomnia. Polysomnographic variables, such as short sleep duration or sleep apnea, did not predict incident chronic insomnia. Mental health, poor sleep, and obesity, but not sleep apnea, are significant risk factors for incident chronic insomnia. Focusing on these more vulnerable groups and addressing the modifiable risk factors may help reduce the incident of chronic insomnia, a common and chronic sleep disorder associated with significant medical and psychiatric morbidity and mortality.


ConvexML: Scalable and accurate inference of single-cell chronograms from CRISPR/Cas9 lineage tracing data.
CRISPR/Cas9 gene editing technology has enabled lineage tracing for thousands of cells in vivo . However, most of the analysis of CRISPR/Cas9 lineage tracing data has so far been limited to the reconstruction of single-cell tree topologies , which depict lineage relationships between cells, but not the amount of time that has passed between ancestral cell states and the present. Time-resolved trees, known as chronograms , would allow one to study the evolutionary dynamics of cell populations at an unprecedented level of resolution. Indeed, time-resolved trees would reveal the timing of events on the tree, the relative fitness of subclones, and the dynamics underlying phenotypic changes in the cell population - among other important applications. In this work, we introduce the first scalable and accurate method to refine any given single-cell tree topology into a single-cell chronogram by estimating its branch lengths. To do this, we leverage a statistical model of CRISPR/Cas9 cutting with missing data, paired with a conservative version of maximum parsimony that reconstructs only the ancestral states that we are confident about. As part of our method, we propose a novel approach to represent and handle missing data - specifically, double-resection events - which greatly simplifies and speeds up branch length estimation without compromising quality. All this leads to a convex maximum likelihood estimation (MLE) problem that can be readily solved in seconds with off-the-shelf convex optimization solvers. To stabilize estimates in low-information regimes, we propose a simple penalized version of MLE using a minimum branch length and pseudocounts. We benchmark our method using simulations and show that it performs well on several tasks, outperforming more naive baselines. Our method, which we name 'ConvexML', is available through the cassiopeia open source Python package.


Genetic basis of ear length in sheep breeds sampled across the region from the Middle East to the Alps.
Ear length in sheep (Ovis aries) shows a wide range of natural variation, from the absence of an outer ear structure (anotia), to small outer ears (microtia), to regular ear length. Up until now, the underlying genetics of this phenotype has been studied in four sheep breeds from China, Jordan and Italy. These studies revealed a broad range of genes significantly associated with ear length, potentially indicating genetic heterogeneity across breeds or geographic regions. In the current study, we performed genome-wide SNP genotyping and haplotype-based mapping, in a population of 340 individuals, to identify loci influencing ear length variation in additional sheep breeds from Slovenia, Croatia, Cyprus and Greece. Additionally, two previously described candidate variants were also genotyped in our mapping population. The mapping model without candidate variant genotypes revealed only one genome-wide significant signal, which was located next to HMX1 on OAR6. This region was previously described as being associated with ear length variation in the Altay and Awassi sheep breeds. The mapping model including the candidate duplication genotype near HMX1 as a fixed effect explained the phenotypic variance on OAR6 and revealed an additional genome-wide significant locus on OAR13 associated with ear length. Our results, combined with published evidence, suggest that a duplication in the evolutionarily conserved region near HMX1 is the major regulator of ear length in sheep breeds descended from a larger region from Central Asia, to the Middle East, Cyprus, Greece and to the Alps. This distribution suggests an ancient origin of the derived allele.


Cartilage formation measured by a novel PIINP assay suggests that IGF-I does not stimulate but maintains cartilage formation ex vivo.
The aim of this study was to investigate the time-dependent effect of insulin-like growth factor-I (IGF)-I on cartilage, evaluated by a novel procollagen type II N-terminal propeptide (PIINP) formation assay. This was performed in a cartilage model. Bovine articular cartilage explants were cultured in Dulbecco's modified Eagle's medium (DMEM):F12 in the presence of 0, 0.01, 0.1, 1, 10, or 100 ng/mL of IGF-I. The viability of the chondrocytes was measured by the colorimetric Alamar blue assay. Collagen formation was assessed from the conditioned medium by the PIINP assay. Proteoglycan levels retained in the explants after 22 days of culture were extracted and measured by the sulfated glycosaminoglycan (sGAG) assay. In the absence of stimulation, PIINP markedly decreased as a function of time (99.4%, p < 0.001). IGF-I dose-dependently stimulated collagen formation and more than 3000% (p < 0.0005) at 100 ng/mL IGF-I at day 20 compared to vehicle control (W/O). IGF-I maintained PIINP at levels comparable to that of day 1. IGF-I dose-dependently protected against proteoglycan loss. IGF-I dose-dependently maintained cartilage formation. The current developed techniques aid the model to represent a more physiologically relevant model to test novel anabolic drugs for osteoarthritis (OA).


Intracellular pH in human resistance arteries in essential hypertension.
To investigate intracellular pH (pHi) in human resistance arteries in essential hypertension, vessels were obtained from small biopsies of skin and subcutaneous fat from 14 untreated patients, and the results were compared with those from 14 matched normotensive control volunteers. Segments of isolated resistance arteries were mounted in a myograph and loaded with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein. Fluorescence signals were monitored using a series of barrier filters and chromatic beam splitters. In this way both resting pHi and the changes in pHi observed during isometric contractions initiated by agonists could be recorded. Resting pHi was not different in vessels from hypertensive patients (hypertensive, 7.24 +/- 0.06 versus control, 7.25 +/- 0.04 pH units). The application of ethylisopropylamiloride (EIPA) and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) demonstrated that both Na(+)-H+ exchange and bicarbonate-dependent membrane mechanisms contributed to pHi homeostasis but that neither system was overactive in hypertension (pHi change with EIPA in vessels from hypertensive versus control subjects was -0.11 +/- 0.02 and 0.13 +/- 0.03 pH units, respectively, and pHi change with DIDS in vessels from hypertensive versus control subjects was -0.097 +/- 0.05 and -0.091 +/- 0.03 pH units, respectively). The application of norepinephrine or 125 mM K+ solution induced contraction in the arterial segments with an accompanying fall in pHi. With norepinephrine this fall was significantly attenuated in vessels from hypertensive patients. These results fail to provide evidence for raised pHi in resistance arteries in human essential hypertension, and contrary to previous reports in circulating blood cells, Na(+)-H+ exchange is not overactive in the vessels of such patients.


Patterns of cervical cancer screening, diagnosis, and follow-up treatment in a state Medicaid fee-for-service population.
Despite being a screening-amenable cancer, cervical cancer is the third most common genital cancer among white women and the most common among African American women. The study objective was to use administrative claims data for CC disease surveillance among recipients enrolled in a state Medicaid fee-for-service (FFS) program. West Virginia (WV) Medicaid FFS administrative claims data for female recipients aged 21-64 years from 2003 to 2008 were used for this study. All medical and prescription claims were aggregated to reflect each recipient's medical care and prescription drug utilization. The yearly prevalence of Pap smear testing declined from 23.9% in 2003 to 15.8% in 2008 in the Medicaid FFS population. During the 6-year study period, persistence with Pap smear testing was low; 41.8% of recipients received no Pap smear testing. Only 73.1% of recipients received Pap smear testing during the year prior to their CC or precancerous cervical lesions (PCL) diagnosis. The likelihood of a CC diagnosis increased with a decrease in Pap smear testing persistence. Only 10.1% of recipients received appropriate follow-up care following a diagnosis of high-grade PCL; only 31.5% of the recipients received appropriate follow-up care for low-grade PCL diagnosis. Although CC preventive services such as screening and PCL follow-up care are covered under Medicaid programs, underutilization of these services by recipients in the Medicaid FFS population is a concern. Results of this study emphasize the need to address disparities in screening and appropriate PCL follow-up care among recipients in the Medicaid FFS population.


Induction of signal transduction pathways related to the pathogenicity of Cryptococcus neoformans in the host environment.
Cryptococcus neoformans, a human pathogenic fungus, infects immunocompromised humans and causes serious diseases such as cerebral meningitis. C. neoformans controls the expression of virulence factors in response to the host environment via various signal transduction pathways. Understanding the molecular mechanisms involved in C. neoformans infection will contribute to the development of methods to prevent and treat C. neoformans-related diseases. C. neoformans produces virulence factors, such as a polysaccharide capsule and melanin, to escape host immunity. Several proteins of C. neoformans are reported to regulate production of the virulence factors. In this review, on the basis of studies using gene-deficient mutants of C. neoformans and animal infection models, we outline the signal transduction pathways involved in the regulation of virulence factors.


Effects of deep wet etching in HF/HNO3 and KOH solutions on the laser damage resistance and surface quality of fused silica optics at 351 nm.
We investigate the interest of deep wet etching with HF/HNO3 or KOH solutions as a final step after polishing to improve fused silica optics laser damage resistance at the wavelength of 351 nm. This comparison is carried out on scratches engineered on high damage threshold polished fused silica optics. We evidence that both KOH and HF/HNO3 solutions are efficient to passivate scratches and thus improve their damage threshold up to the level of the polished surface. The effect of these wet etchings on surface roughness and aspect is also studied. We show that KOH solution exhibit better overall surface quality that HF/HNO3 solution in the tested conditions. Given the safety difficulties associated with the processing with HF, KOH solution appears as a pertinent alternative to HF deep wet etching.


Loss Function Based Ranking in Two-Stage, Hierarchical Models.
Performance evaluations of health services providers burgeons. Similarly, analyzing spatially related health information, ranking teachers and schools, and identification of differentially expressed genes are increasing in prevalence and importance. Goals include valid and efficient ranking of units for profiling and league tables, identification of excellent and poor performers, the most differentially expressed genes, and determining "exceedances" (how many and which unit-specific true parameters exceed a threshold). These data and inferential goals require a hierarchical, Bayesian model that accounts for nesting relations and identifies both population values and random effects for unit-specific parameters. Furthermore, the Bayesian approach coupled with optimizing a loss function provides a framework for computing non-standard inferences such as ranks and histograms.Estimated ranks that minimize Squared Error Loss (SEL) between the true and estimated ranks have been investigated. The posterior mean ranks minimize SEL and are "general purpose," relevant to a broad spectrum of ranking goals. However, other loss functions and optimizing ranks that are tuned to application-specific goals require identification and evaluation. For example, when the goal is to identify the relatively good (e.g., in the upper 10%) or relatively poor performers, a loss function that penalizes classification errors produces estimates that minimize the error rate. We construct loss functions that address this and other goals, developing a unified framework that facilitates generating candidate estimates, comparing approaches and producing data analytic performance summaries. We compare performance for a fully parametric, hierarchical model with Gaussian sampling distribution under Gaussian and a mixture of Gaussians prior distributions. We illustrate approaches via analysis of standardized mortality ratio data from the United States Renal Data System.Results show that SEL-optimal ranks perform well over a broad class of loss functions but can be improved upon when classifying units above or below a percentile cut-point. Importantly, even optimal rank estimates can perform poorly in many real-world settings; therefore, data-analytic performance summaries should always be reported.


Identification and analysis of knee-ankle-foot orthosis design requirements based on a feedback survey of orthosis users in India.
The world is advancing towards a technological revolution in various fields, yet the assistive devices available for people with disability, especially in developing countries, are in the most primitive stage. For many years, lower limb orthotics has been a neglected area of research and there is an urgent need to address the problems faced by lower limb orthosis users to enable them to lead an independent life. This work is a first step in this direction and aims to identify and analyse the needs of knee-ankle-foot orthosis (KAFO) users in India. A structured feedback survey of 29 KAFO users was conducted at three rehabilitation centres located in South India. A feedback questionnaire and a novel outcome measure tool (trigger cards) were used as means to assess user satisfaction about their existing KAFOs. The results of the survey were analysed to obtain quantitative and qualitative outcomes. The survey identifies various biomechanical and functional issues associated with lower limb orthosis design. The results of the survey imply that there is an urgent need to solve issues, especially related to locked orthotic knee joint design. Additionally, it sheds light on the lifestyle and socio-economic issues of KAFO users that are likely prevalent in many other low- and middle-income countries. The outcomes of this survey can motivate and guide researchers to design improved orthotic solutions to meet the needs of lower limb orthosis users all over the world. Implications for Rehabilitation • This is a first of its kind survey that brings forth the needs of lower limb orthosis users in India, and is an important step towards rehabilitation and empowerment of people with lower limb disability. • The pilot survey helps to identify critical areas for design improvements in a knee-ankle-foot orthosis. • The outcomes of this survey can help researchers to design functionally improved assistive devices that better meet the needs of users than currently available technology in developing countries such as India.


[Effect of xinmaitong on wild-type p53 gene expression in rabbits with carotid endothelial injury].
To explore the mechanism of prevention and treatment of post-PTCA restenosis with Xinmaitong (XMT). Rabbit carotid endothelial injury model was established using Fishman air drying method. Effect of XMT on model rabbits wild-type p53 gene expression was observed by tissue in situ hybridization. p53 gene expression appeared on the 3 days after operation, enhanced on the 7 days, reached the peak on the 14th day, weakened on the 21th, and still showed on the 28th day. The strongest expression was shown in rabbits treated by XMT, second in those treated by Warfarin, and the weakest in the operated control group. XMT could promote the high expression of wild-type p53 gene expression in rabbit with carotid endothelial injury, which is possibly one of the important mechanism of XMT in preventing and treating arterial restenosis.


Analyses and treatments of postoperative nasal complications after endonasal transsphenoidal resection of pituitary neoplasms.
In this study, we analyze and discuss the treatments of postoperative nasal complications after endonasal transsphenoidal resection of pituitary neoplasms (PNs). We performed 129 endonasal transsphenoidal resections of PNs and analyzed and treated cases with nasal complications. After endonasal transsphenoidal resection of PNs, there were 26 cases of postoperative nasal complications (20.1%), including nasal hemorrhage (4.8%), cerebrospinal fluid rhinorrhea (6.9%), sphenoid sinusitis (2.3%), atrophic rhinitis (1.6%), olfactory disorder (1.6%), perforation of nasal septum (0.8%), and nasal adhesion (2.3%). All patients clinically recovered after therapy, which included treatment of the cavity through nasal endoscopy, intranasal corticosteroids, and nasal irrigation. We propose that regular nasal endoscopic review, specific nasal medications, and regular nasal irrigation can effectively clear nasal mucosal hyperemia-induced edema and nasal/nasoantral secretions, as well as promote regeneration of nasal mucosa, prevent nasal adhesion, maintain the sinus cavity drainage, and accelerate the recovery of the physiological function of the paranasal sinus. Timely treatment of patients with nasal complications after endonasal transsphenoidal resections of PNs could greatly relieve the clinical symptoms. Nasal cleaning is very beneficial to patients after surgery recovery.


Myosin isozyme synthesis and mRNA levels in pressure-overloaded rabbit hearts.
The in vivo synthesis rates of myosin isozyme heavy chains beta and alpha were measured in right ventricular (RV) muscle at 2 and 4 days following pulmonary artery constriction in rabbits, together with measurements of their relative mRNA levels. The synthesis rate of beta-myosin heavy chains was elevated in 2-day (0.27 +/- 0.06 day-1 or 2.5 +/- 0.7 mg/g RV/day, mean +/- SD) and in 4-day (0.25 +/- 0.08 day-1 or 2.8 +/- 1.0 mg/g RV/day) pressure overload, when compared to untreated rabbits (0.15 +/- 0.04 day-1 or 1.5 +/- 0.4 mg/g RV/day). However, the synthesis rates of alpha-myosin heavy chains in the same hearts were not altered significantly. There was a differential increase in the fractional synthesis rate of beta vs. alpha heavy chains in 2-day and 4-day pressure overload and in 2-day shams, suggesting switching toward beta heavy chain synthesis had occurred at these time points. beta heavy chain synthesis, as a proportion of total (alpha + beta) heavy chain synthesis, was significantly higher in 4-day pressure overload (78 +/- 9%) than in 4-day sham rabbits (63 +/- 6%). This increase in relative beta-synthesis was associated with a significant increase in the relative proportion of beta heavy chain mRNA level (76 +/- 13% vs. 56 +/- 7%). Furthermore, relative beta-synthesis and the beta-mRNA levels correlated linearly with each other in all experimental groups.(ABSTRACT TRUNCATED AT 250 WORDS)


A folate-rich diet is as effective as folic acid from supplements in decreasing plasma homocysteine concentrations.
Background & Aims: At least 500 mug of folic acid are required daily to treat hyperhomocysteinemia. To reach this amount by dietary changes alone may be difficult because food has a low folic acid content and bioavailability. No studies have compared the effects of similar amounts of additional folate derived from a combination of folate-rich and fortified foods or folic acid from supplements on plasma total homocysteine (tHcy) concentrations, which was the aim of this study. Methods: Twenty male patients with hyperhomocysteinemia and coronary artery disease were included in a randomized, crossover intervention trial. Patients were treated daily with a combination of foods containing approximately 500 mug of folate or with one 500 mug capsule of synthetic folic acid over two five-week periods separated by a five-week wash-out period. Results: Plasma folate increased markedly (p<0.001) and plasma tHcy decreased (p<0.001) with both therapies. Folate-rich foods decreased tHcy by 8.6% (95% CI: -15.9 to -1.2) and synthetic folic acid capsules by 8% (95% CI: -13.3 to -2.7). Conclusions: This study shows, for the first time in the literature, that a folate-rich diet is as effective as folic acid capsules in decreasing plasma tHcy concentrations and adds further support to the recommendation of those diets to prevent cardiovascular disease.


Dermatological features in Behçet disease-associated vena cava obstruction.
Behçet disease (BD) is a multisystemic disorder that is classified as vasculitis and can affect all types and sizes of blood vessels. Although vascular diseases are not regarded as essential for a diagnosis of BD, vascular complications can be fatal. The purpose of this study was to examine the clinical characteristics of 14 patients with BD accompanied by an obstruction of the superior vena cava (VC), the inferior VC, or both, and then determine the dermatological features that indirectly suggest VC obstructions in patients with BD. Among the 3500 patients registered at the BD Specialty Clinic of Severance Hospital from 1997 to 2006, 14 patients were diagnosed with BD-associated VC obstructions. Their medical records were reviewed. Nine of the 14 patients with BD were male, and the age range for disease onset was 11-46 years. Computed tomography revealed superior VC thrombosis in seven patients, inferior VC thrombosis in four patients, and the involvement of both in three patients. The most prominent features in superior VC obstruction were facial oedema (n = 7, 50%) and neck vein distension (n = 6, 43%). Other features included dilated veins in the chest wall, upper body oedema, proptosis, and upper extremity swelling. Patients with an inferior VC obstruction commonly had dilated veins on the abdominal walls. These patients also had abdominal distension and lower extremity swelling. Corticosteroids, colchicine, immunosuppressants, and anticoagulants with or without heparinization were the most common treatment regimens giving good response. Attempts to treat patients with a high dose of steroid pulse therapy were ineffective. Ballooning venoplasty was a useful therapeutic modality for VC thrombosis during the early stage, although this procedure could not be performed in patients with BD with severe thrombosis. Several cutaneous features of VC obstruction should be recognized in patients with BD. Early detection might permit an early diagnosis and treatment of occlusion of the VC.


Social disadvantages in childhood and risk of all-cause death and cardiovascular disease in later life: a comparison of historical and retrospective childhood information.
Childhood socioeconomic circumstances have been shown to contribute to adult mortality. The purpose of this study was to compare the association between objective historical records and recalled questionnaire-based information on childhood socioeconomic position (SEP) with regard to cardiovascular and all-cause mortality. We examined the association between a socially disadvantaged childhood and all-cause mortality, cardiovascular disease (CVD) mortality, coronary heart disease (CHD) mortality, and acute coronary events among male participants in the Kuopio Ischemic Heart Disease (KIHD) Risk Factor Study, a population-based cohort study in eastern Finland with follow-up until 2002. The historical data on childhood factors were collected from school health records (n = 698), mainly from the 1930s to the 1950s. Recall data on socioeconomic conditions in childhood were obtained from the baseline examinations of the KIHD cohort (n = 2,682) in 1984-89. According to original school health records the men who were socially disadvantaged in childhood had a 1.41-fold (95% confidence interval 1.01-1.97) age-adjusted and examination-year-adjusted risk of all-cause death, a 1.32-fold (0.83-2.11) risk of CVD death, a 1.48-fold (0.85-2.57) risk of CHD death, and a 1.50-fold (1.02-2.20) risk of acute coronary events. After adjustment for biological and behavioural risk factors and for the SEP in adulthood the association was attenuated in all-cause death but did not change in CVD death, CHD death, and acute coronary events. On the contrary, the questionnaire-based recalled childhood data on childhood SEP showed no associations with mortality or acute coronary events. With regard to adult mortality, the use of historical records concerning hygiene and living conditions collected in childhood may either provide more accurate measures of early-life socioeconomic conditions or capture more relevant aspects of childhood socioeconomic disadvantage than retrospective recall data.


Microbial synthesis of pyrroloquinoline quinone.
Pyrroloquinoline quinone (PQQ) is a peptide-modified natural product. PQQ has important physiological functions such as anti-oxidation, anti-aging, and immunity enhancement. However, due to the lack of in-depth understanding of PQQ biosynthesis and regulation, inefficient PQQ production level limits its wide application. Accordingly, there is still an urgent need to develop high-yielding strains for synthesis of PQQ. This paper reviewed the research and development trends on the PQQ biosynthetic pathways, catalytic reaction mechanism of key enzymes, and the selection of high-yielding strains, which also prospects for the future construction of PQQ biosynthetic microbial cell factories.


The Relation Between Tobacco Tax Structure and Corruption in European Union Member States.
Background: Taxing tobacco products is one of the most effective tobacco control measures, and most countries apply a combination of specific taxes, which comprise a fixed amount per cigarette or gram of hand-rolling tobacco, and ad valorem taxes, which increase in proportion to the cost of the product. Since specific taxes reduce price differentials across tobacco product ranges while ad valorem taxes amplify them, we hypothesised that tobacco companies seeking to minimise the effect of tax increases on sales across a range of products will tend to favour, and hence lobby for, ad valorem rather than specific taxes; and that relatively corrupt governments would be more susceptible to such lobbying and hence, more likely to favour ad valorem taxes. Methods: We searched for cigarette tax data and Transparency International Corruption Perceptions Index (CPI) scores for current 28 EU Member States for the years 1995 to 2017/8. Trends in cigarette tax levels and the ratio of ad valorem to specific taxes at a national and mean EU level were analysed by visual inspection, the within-country relation between the ad valorem to specific tax ratio and CPI scores over time by time-series regression analysis, and at EU level, for which complete data were available from 1995 to 2017, using a multi-level regression model. Results: Within most Member States, the ad valorem to specific cigarette tax ratio declined over the study period and was not significantly associated with corruption score. However, at an aggregate EU-level, our multi-level model indicated that reduced corruption was associated with a significant increase in the ad valorem to specific cigarette tax ratio, by 0.04 (95% confidence interval: 0.003-0.077, p < 0.036) per unit increase in CPI score. Conclusions: The ratio of ad valorem to specific taxes declined in most EU Member States over the study period, with no evidence that those with higher levels of perceived corruption tended to favour ad valorem taxes.


High-grade squamous intraepithelial lesion in women aged <30 years has a prevalence pattern resembling low-grade squamous intraepithelial lesion.
Cervical cytology is the cervical cancer screening test for women aged <30 years because of the low specificity of human papillomavirus tests in this age group. The Bethesda System classifies cervical intraepithelial neoplasia grade 2 (CIN 2) and grade 3 (CIN 3) as high-grade intraepithelial lesions (HSIL). In this study, the authors subclassified cytologic HSIL as suggestive of CIN 2 (HSIL-CIN 2) or CIN 3 (HSIL-CIN 3) and evaluated whether there was a correlation between these findings and age for screened and unscreened women. The study included 2,002,472 cervical smears collected from women who had at least 1 previous test (screened) and 217,826 previously untested women (unscreened). The laboratory has been using the Bethesda System since 1998 with the subcategorization of HSIL-CIN 2 and HSIL-CIN 3. For unscreened women, the prevalence of low-grade intraepithelial lesion (LSIL) and HSIL-CIN 2 decreased with age, whereas the prevalence of HSIL-CIN 3 increased. The prevalence of HSIL-CIN 2 was greater than that of HSIL-CIN 3 for women up to age 29 years (prevalence ratio [PR], 4.73; 95% confidence interval [CI], 3.90-5.75) and lower for the groups ages 30 to 49 years (PR, 0.66; 95% CI, 0.50-0.87) and ≥ 50 years (PR, 0.21; 95% CI, 0.12-0.36). For screened women, the prevalence of HSIL-CIN 2 also was greater in the group aged ≤ 29 years (PR, 2.72; 95% CI, 2.49-2.97). The prevalence pattern of HSIL suggestive of CIN 2 resembled the pattern observed in LSIL and was more prevalent than HSIL suggestive of CIN 3 in younger women. The impact of screening was less evident when HSIL was suggestive of CIN 2. A conservative approach for younger women who have HSIL is important for management guidance.


Pattern of expression of the jun family of transcription factors during the early development of the inner ear: implications in apoptosis.
Jun transcription factors have been implicated in the regulation of cell proliferation, differentiation and apoptosis. We have investigated the relationship between Jun expression and cell death in the developing chicken inner ear. c-jun and junD transcripts were expressed in the epithelium of the otic placode and otic vesicle. c-jun expression was restricted to the dorsal area of the otic pit (stages 14-17), dorsal otic vesicle and cochleo-vestibular ganglion (stages 18-20). junD expression was transient and occurred in the dorsal and upper medial aspects of the otic pit and otic cup, but it was down-regulated in the otic vesicle. A parallel TUNEL analysis revealed that expression of c-jun co-located within areas of intense apoptosis. Furthermore, phosphorylation of c-Jun at serine-63 by Jun amino-terminal-kinases was detected in the dorsal otic pit, otic vesicle and cochleo-vestibular ganglion. c-Jun protein exhibited DNA binding activity, as assessed by gel mobility shift assays. The association between c-Jun and apoptosis was further demonstrated by studying nerve growth factor-induced apoptosis in cultured otic vesicles. Nerve growth factor-induced cell death and c-Jun phosphorylation that were suppressed by insulin-like growth factor-I and by viral-mediated overexpression of Raf, which had survival effects. In conclusion, the precise regulation of the expression and activity of Jun proteins in the otic primordium suggests that it may operate as a fundamental mechanism during organogenesis.


Socioeconomic inequalities in mental health and health-related quality of life (HRQOL) in children and adolescents from 11 European countries.
To assess the presence and magnitude of social inequalities in mental health and health-related quality of life (HRQOL) in the population aged 8-18 years in 11 European countries. Cross-sectional surveys were carried out in representative samples of children/adolescents (8-18 years) from the participating countries of the KIDSCREEN project. Mental health was assessed using the Strengths and Difficulties Questionnaire (SDQ), and HRQOL by means of the KIDSCREEN-10. Socioeconomic status (SES) was assessed using the Family Affluence Scale and parental level of education. The association between health outcomes and SES was analyzed with the regression-based relative index of inequalities (RII) and population attributable risk. A total of 16,210 parent-child pairs were included. The SDQ showed inequalities in mental health according to family level of education in all countries (RII = 1.45; 1.37-1.53). The RII for HRQOL was 2.15 (1.79-2.59) in the whole sample, with less consistent results by age and country. Socioeconomic inequalities in mental health were consistently found across Europe. Future research should clarify the causes of these inequalities and define initiatives which prevent them continuing into adulthood.


ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma.
ATF3 was a transcription factor involved in the progression of certain cancers. Here, we sought to explore the expression and biological function of ATF3 in esophageal squamous cell carcinomas (ESCC). The prognostic significance of ATF3 expression was evaluated in 150 ESCC samples and 21 normal squamous cell epithelium tissues. Results showed that ATF3 was down-regulated in ESCC lesions compared with paired non-cancerous tissues and low tumorous ATF3 expression significantly correlated with shorter overall survival (OS) and disease-free survival (DFS). Cox regression analysis confirmed that ATF3 expression was an independent prognostic factor. Experimentally, forced expression of ATF3 led to decreased growth and invasion properties of ESCC cells in vitro and in vivo, whereas knockdown of ATF3 did the opposite. Furthermore, ATF3 upregulated the expression of MDM2 by increasing the nuclear translocation of P53 and formed an ATF3/MDM2/MMP-2 complex that facilitated MMP-2 degradation, which subsequently led to inhibition of cell invasion. Finally, we showed that Cisplatin could restrain the invasion of ESCC cells by inducing the expression of ATF3 via P53 signaling. Combined, our findings highlight a suppressed role for ATF3 in ESCC and targeting ATF3 might be a potential therapeutic strategy.


From spin glass to quantum spin liquid ground states in molybdate pyrochlores.
We present new magnetic heat capacity and neutron scattering results for two magnetically frustrated molybdate pyrochlores: S=1 oxide Lu_{2}Mo_{2}O_{7} and S=1/2 oxynitride Lu_{2}Mo_{2}O_{5}N_{2}. Lu_{2}Mo_{2}O_{7} undergoes a transition to an unconventional spin glass ground state at T_{f}∼16 K. However, the preparation of the corresponding oxynitride tunes the nature of the ground state from spin glass to quantum spin liquid. The comparison of the static and dynamic spin correlations within the oxide and oxynitride phases presented here reveals the crucial role played by quantum fluctuations in the selection of a ground state. Furthermore, we estimate an upper limit for a gap in the spin excitation spectrum of the quantum spin liquid state of the oxynitride of Δ∼0.05 meV or Δ/|θ|∼0.004, in units of its antiferromagnetic Weiss constant θ∼-121 K.


COVID-19: Current challenges regarding medical healthcare supplies and their implications on the global additive manufacturing industry.
The covid-19 outbreak has caused a shortage of masks and other healthcare products for the general public around the globe. In addition, it has also affected the supply of personal protective equipment (PPE) used by healthcare services because of a sudden increase in their demand. This significant disruption in the global supply chain of these products resulted in, leaving many staff and patients without protection. The additive manufacturing (AM) industry is going through extraordinary times and can provide emergency responses to help deal with the global crisis caused by the COVID-19 pandemic. The objective of the present work is therefore to perform an up-to-date review to determine the capacity of AM to provide exclusive benefits for the medical healthcare supplies sector to fight this current situation. In this review, it is found that AM technology has proved that it can be used as a volume manufacturing technology for the ongoing crisis. However, the standardization and certification are appeared to represent the main challenges for adopting the AM in healthcare against COVID-19. Furthermore, additively manufactured materials for medical applications must be developed for medical environments. Most printed medical products for COVID-19 require biocompatibility evaluation and shall prove their ability to sterilize. Finally, this review concluded that AM technology can fulfill the requirements of face masks and ventilator parts for healthcare systems for proper controlling and treating of COVID-19 patients when the safety and efficacy of these devices are ensured.


The PH-20 protein in cynomolgus macaque spermatozoa: identification of two different forms exhibiting hyaluronidase activity.
In these experiments, we have characterized the bifunctional sperm protein PH-20 in macaque sperm and studied its hyaluronidase activity. Intact sperm were evaluated before the acrosome reaction (AR), and a soluble form of PH-20 released during acrosomal exocytosis was also investigated. Western blots of SDS-PAGE of acrosome-intact sperm extracts revealed a 64-kDa form of PH-20 was recognized by a polyclonal antibody (R-10) raised in rabbits against purified, recombinant cynomolgus macaque sperm PH-20. The soluble components released during the AR which were recognized by the R-10 antibody included both the 64-kDa form and a 53-kDa form of PH-20. An ELISA-like procedure for determining PH-20 hyaluronidase activity indicated that acrosome-intact sperm exhibited two peaks of hyaluronidase activity near pH 4 and > or = pH 7. The majority of enzyme activity in acrosome-intact sperm extracts occurred at neutral pH, while the soluble hyaluronidase activity released at the AR was predominantly acid-active. Hyaluronidase activity of PH-20 at different pH optima was investigated using hyaluronic acid substrate gel electrophoresis, and results indicated that the 64-kDa polypeptide had a broad range, with the majority of activity at neutral pH (pH 7). The 53-kDa polypeptide in sperm extracts only exhibited activity at acid pH (pH 4). The hyaluronidase activities of both enzymes could be inhibited by apigenin. The soluble PH-20 hyaluronidase activity released during the AR was primarily of the acid-active 53-kDa form. Fine structural localization of PH-20 using Fab fragments of R-10 IgG demonstrated that PH-20 was associated not only with sperm membranes, but also with the dispersing acrosomal contents. These data suggest that the more neutral-active form of PH-20 (64 kDa) is present on the plasma and inner acrosomal membranes and gives rise to the soluble acid-active form at the time of the AR. The generation of the soluble form of PH-20 may result from the action of acrosomal enzymes, which could include proteases, glycosidases, and phospholipases.


Algal swimming velocities signal fatty acid accumulation.
The use of microalgae for biofuel production will be beneficial to society if we can produce biofuels at large scales with minimal mechanical energy input in the production process. Understanding micro-algal physiological responses under variable environmental conditions in bioreactors is essential for the optimization of biofuel production. We demonstrate that measuring micro-algal swimming speed provides information on culture health and total fatty acid accumulation. Three strains of Chlamydomonas reinhardtii were grown heterotrophically on acetate and subjected to various levels of nitrogen starvation. Other nutrient levels were explored to determine their effect on micro-algal kinetics. Swimming velocities were measured with two-dimensional micro-particle tracking velocimetry. The results show an inverse linear relationship between normalized total fatty acid mass versus swimming speed of micro-algal cells. Analysis of RNA sequencing data confirms these results by demonstrating that the biological processes of cell motion and the generation of energy precursors are significantly down-regulated. Experiments demonstrate that changes in nutrient concentration in the surrounding media also affect swimming speed. The findings have the potential for the in situ and indirect assessment of lipid content by measuring micro-algal swimming kinetics.


Host factors predisposing to anaerobic infections.
Anaerobic bacterial infections of man are typically associated with locally comprised tissues and commonly also with a systemically compromised host. Thus, any condition that interferes with the systemic defence mechanisms of the host and thus predisposes to infection in general, increases the incidence of anaerobic infections also. It is generally true to say that anaerobic infections occur secondary to mechanical disruption of anatomical barriers, usually percutaneous in clostridial infections; in nonclostrial anaerobic infections the disrupted barriers are commonly mucosal surfaces where the offending organisms exist as the dominant part of normal bacterial floras. The most important single factor which enables anaerobes to flourish in the tissues is low oxygen tension which is usually the result in the first instance of a compromised blood supply and poor tissue perfusion. A variety of other predisposing factors may operate in concert, enabling the nonsporing anaerobes to cause a diversity of infective states in many different parts of the body. These host determinants are considered in relation to clostridial, nonclostridial and postsurgical anaerobic sepsis.


[Possibilities of immunotherapy in malignant melanoma].
The immune response of malignant melanoma bases on evident humoral and cellular defence mechanisms of the host against his tumor. Of special interest is a defective immune response, developing in the course of the disease. Hence, any immunotherapy aims at an immunostimulation and immunoregulation. Local and systemic nonspecific immunotherapy try to raise an immune response against the tumor by stimulating unspecifically the whole immune system. On the contrary, specific immunotherapy tries to stimulate directly the defence mechanisms against the tumor by transfusion of antisera (passive immunotherapy) and sensitized cells (adoptive immunotherapy) and by immunizing the patient with tumor tissue (active immunotherapy). One of the best ways in therapy of melanoma seems to be the combination of immunotherapy with chemotherapy, as yet employed in BCG and DTIC treatment.


Physician-assisted suicide and euthanasia in Washington State. Patient requests and physician responses.
To estimate how often physicians receive requests for physician-assisted suicide and euthanasia and to describe a case series of patient requests for physician-assisted suicide and euthanasia, including physician responses to these requests. A mailed, anonymous two-part questionnaire. A total of 828 physicians returned questionnaires sent to 1453 potential respondents, for a response rate of 57%. Questionnaires were mailed to random sample (25%) of primary care physicians and all physicians in selected medical subspecialties in Washington State. The frequency of explicit patient requests for physician-assisted suicide and euthanasia reported by physicians and individual case descriptions of patient characteristics, physician perceptions of patient concerns, and physician responses to patient requests. In the past year, 12% of responding physicians received one or more explicit requests for physician-assisted suicide, and 4% received one or more requests for euthanasia. These physicians provided 207 cases descriptions. The diagnoses most often associated with requests were cancer, neurological disease, and the acquired immunodeficiency syndrome (AIDS). The patient concerns most often perceived by physicians were worries about loss of control, being a burden, being dependent on others for personal care, and loss of dignity. Physicians provided assistance more often to patients with physical symptoms. Physicians infrequently sought advice from colleagues. Of 156 patients who requested physician-assisted suicide, 38 (24%) received prescriptions, and 21 of these died as a result. Of 58 patients who requested euthanasia, 14 (24%) received parenteral medication and died. Patient request for physician-assisted suicide and euthanasia are not rare. As perceived by physicians, the most common patient concerns at the time these requests are made are nonphysical. Physicians occasionally provide these practices, even though they are currently illegal in Washington State. Physicians do not consult colleagues often about these requests. These findings raise the question of how to ensure quality in the evaluation of patient requests for physician-assisted death.